The show premiered on March 28, 2005 as a replacement for The Screen Savers. The show originally aired live five days a week; the week of January 2, 2006, the show was truncated to air for only four days a week. Fridays featured a "Mash Up" that compiled segments from the week's four live shows. The show returned to live shows five days a week in September 2008, but G4 announced as of March 2, 2009, Attack of the Show! would go to four days a week with a "best of" show running in the Friday time-slot. Attack of the Show! aired its 1,000th episode on July 9, 2009. Attack of the Show! upgraded to High Definition on April 5, 2010 to coincide with its first ever "Epic April" event of new premieres and stunts. Along with the addition of Candace Bailey, on January 11, 2011 Attack of the Show "respawned" with new graphics and a new set.
Host Kevin Rose left the G4 Network and AOTS on May 22, 2005. The show announced a search for a new host among the show's audience, and for several months various candidates auditioned on the air. In the end none of the candidates were actually selected, although this was never actually announced on the air. Sarah Lane, who had previously been a co-host and segment contributor, gradually became the show's official host alongside Kevin Pereira. She continued in this capacity until April 6, 2006, when she and Brendan Moran announced their engagement and stated that they would be leaving the show. The Morans appeared on the show, via satellite, on July 13, 2006, to talk about their adventures.
Olivia Munn was hired to replace Lane and began hosting with Kevin Pereira on Monday, April 10, 2006. Zach Selwyn, the runner up on ESPN's Dream Job, was hired to replace Moran and started his hosting duties a week later on April 17, 2006. Munn said at the 2010 Comic Con that she would not be leaving AOTS, though she only occasionally appeared on the show for the rest of that year. She was still featured in the opening credits and other AOTS set pieces and transitions until December 6, 2010.
After the departure of Munn, AOTS featured several guest co-hosts including: Candace Bailey, Jessica Chobot, Alison Haislip, Carrie Keagan, Alessandra Torresani, Sara Underwood, and Morgan Webb. Bailey became the new co-host with Pereira on January 11, 2011. AOTS has also featured Chris Hardwick, Chris Gore, and Blair Herter in the occasional absence of Kevin Pereira. In addition to the interim cohosts, Grace Helbig has also filled in for Candace Bailey.
On May 9, 2012, Kevin Pereira announced during Attack Of The Show that he is officially leaving AOTS to pursue work in television production. As he stated; “I plan to turn my attention to full-time with my production company Super Creative and live touring nerd-carnival leetUP," His last day on AOTS was May 31. Beginning August 28, 2012, the show debuted a new opening credit, with no reference to permanent or guest hosts. On October 26, 2012 G4 announced that they would end the show at the end of the year.